Highly enriched cardiomyocytes from human embryonic stem cells

Xu, Xiuqin; Zweigerdt, Robert; Soo, Set Yen; Ngoh, Z.X.; Tham, Su Chin; Wang, S.T.; Graichen, Ralph; Davidson, Bruce P.; Colman, Alan; Sun, William

doi:10.1080/14653240802105307

Cited by 100 publications

(71 citation statements)

References 41 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…In addition, we found that the cardiac muscle phenotype of the P19-derived cardiomyocytes was maintained when growing on collagen, a commonly used natural material in tissue engineering A similar cardiac muscle cell enrichment strategy has been employed using human and mouse ESCs, in which a MHC promoter drove antibiotic resistance or GFP expression [39][40][41][42][43][44][45][46][47]. A previous publication reported an 99% purity for human ESC-derived cardiomyocytes used the -MHC promoter driving neomycin resistance approach [44]. The higher enrichment in these ES studies may be due to differences in the promoter specificity due to an integration site in the cells such as near strong lineage-dependent enhancers, the nature of the construct, or simply differences in the method of assessing enrichment.…”

Section: Discussionmentioning

confidence: 99%

A P19 Cardiac Cell Line as a Model for Evaluating Cardiac Tissue Engineering Biomaterials

Dawson¹,

Boisvenue²,

Skerjanc³

et al. 2009

TOTERMJ

View full text Add to dashboard Cite

Abstract:Our objective was to develop a suitable cardiomyocyte progenitor cell line for use in testing biomaterials as potential scaffolds in cardiac tissue engineering. We transfected P19 cells with the human cardiac -actin promoter driving the gene for puromycin resistance, to create a stable cardiomyocyte-selectable P19 cell line, termed P19(CAPuro). Puromycin selection resulted in a 4-10 fold enrichment of cardiac muscle gene expression and a 3-fold enrichment in cardiomyocytes. Morphological, biochemical, and functional analyses were used to evaluate the properties of P19(CAPuro) cardiomyocytes in the presence and absence of a novel cross-linked collagen-based biomaterial. The collagen-based biomaterial was able to support appropriate viability, gene expression, and cardiomyocyte function. Therefore, P19(CAPuro) cells are suitable for examining biomaterials as potential scaffolds and this approach could be used for rapidly screening biomaterials for designing future human embryonic stem cell therapies.

show abstract

Section: Discussionmentioning

confidence: 99%

A P19 Cardiac Cell Line as a Model for Evaluating Cardiac Tissue Engineering Biomaterials

Dawson¹,

Boisvenue²,

Skerjanc³

et al. 2009

TOTERMJ

View full text Add to dashboard Cite

show abstract

“…hiPSCs (clone hCBiPSC2) 1 were cotransfected with the bicistronic vector pCAG_rNIS_IRES2_Venus_nucmem (Figure 1A) and the plasmid ␣MHCneoPGKhygro, 26 enabling selection of hygromycin-resistant transgenic cell clones. Six independent Venus pos clones (designated rNIS clones 1, 2, 4, 5, 6, 7) were analyzed for stability of transgene expression.…”

Section: Functional Nis Expression In Transgenic Hipsc Clonesmentioning

confidence: 99%

Transplantation and Tracking of Human-Induced Pluripotent Stem Cells in a Pig Model of Myocardial Infarction

et al. 2012

View full text Add to dashboard Cite

Background-Evaluation of novel cellular therapies in large-animal models and patients is currently hampered by the lack of imaging approaches that allow for long-term monitoring of viable transplanted cells. In this study, sodium iodide symporter (NIS) transgene imaging was evaluated as an approach to follow in vivo survival, engraftment, and distribution of human-induced pluripotent stem cell (hiPSC) derivatives in a pig model of myocardial infarction. Methods and Results-Transgenic hiPSC lines stably expressing a fluorescent reporter and NIS (NIS pos -hiPSCs) were established. Iodide uptake, efflux, and viability of NIS pos -hiPSCs were assessed in vitro. Ten (Ϯ2) days after induction of myocardial infarction by transient occlusion of the left anterior descending artery, catheter-based intramyocardial injection of NIS pos -hiPSCs guided by 3-dimensional NOGA mapping was performed. Dual-isotope single photon emission computed tomographic/computed tomographic imaging was applied with the use of 123 I to follow donor cell survival and distribution and with the use of 99m TC-tetrofosmin for perfusion imaging. In vitro, iodide uptake in NIS pos -hiPSCs was increased 100-fold above that of nontransgenic controls. In vivo, viable NIS pos -hiPSCs could be visualized for up to 15 weeks. Immunohistochemistry demonstrated that hiPSC-derived endothelial cells contributed to vascularization. Up to 12 to 15 weeks after transplantation, no teratomas were detected. S tem cell-based therapies are being actively explored as a potentially innovative therapeutic strategy for various genetic and acquired diseases. Recently, the possibility of reprogramming human somatic cells into human-induced pluripotent stem cells (hiPSCs) that are able to differentiate into all cell lineages present in the heart 1-4 has opened novel opportunities for myocardial repair. With respect to the potential therapeutic application of pluripotent stem cell derivatives, major progress Conclusions-This Editorial see p 388 Clinical Perspective on p 439However, there are still major hurdles and risks to overcome with regard to PSC-based heart repair. These include safety risks, especially the potential of teratoma and tumor formation, 2 low cell retention and engraftment rates, 8 -11 and the general question of whether engraftment of hiPSCs after simple intramyocardial cell injection leads to formation of functional tissue, such as de novo vasculature or myocardium, and results in significant clinical benefits. 12 Although some of these issues can be addressed in vitro or in appropriate small-animal models, others will require exploration in large-animal models, which are more similar to humans. 13 Transplanted human cardiomyocytes, for example, are unlikely to fully functionally integrate with rodent myocardium because of highly dissimilar beating rates. 11 Therefore, meaningful assessment of human cells for heart repair must be demonstrated in large-animal models such as dogs, pigs, or monkeys. 13 Clearly, advanced imaging technologies allowing for l...

show abstract

“…A diversity of cardiac myocytes are generated in EB cultures [42], and commitment to cardiac fate can be achieved in defined media [43] and enhanced by co-culture with inducing stromal elements, such as the end2 endoderm-like cell line [44]. Similar to the lineage-tracing experiments, a useful approach has been to use a cardiac reporter (for example a cardiac-specific promoter regulating expression of GFP) to select for those cells that commit to cardiac fate [45][46][47]. However, this can also be achieved by modeling the normal developmental signals thought to direct cardiac fate [27,48] or using chemically defined media [28,49] so that highly enriched cardiomyocyte populations can be purified from density gradients.…”

Section: Human Es-derived Cardiomyocytes and Ipsmentioning

confidence: 99%

Embryonic stem cells as a model for cardiac development and disease

Evans

2008

Drug Discovery Today: Disease Models

View full text Add to dashboard Cite

SummaryThis review highlights recent progress in the use of embryonic stem cell (ESC) systems for studying and treating cardiovascular disease. Although ESCs represent an in vitro system, they can provide a rich source of progenitor cells, and this has been exploited recently to identify novel precursors and to investigate the lineage relationships among various cell types that comprise the developing heart, including cardiac muscle, endothelium, and smooth muscle. ESCs grown in aggregates (embryoid bodies) recapitulate normal developmental programs. Since they can be grown under defined culture conditions, they have been used to systematically identify specific genes and signaling pathways that promote cardiogenesis. A major goal is to optimize the production of cardiac progenitors and differentiated cell types, and to test their ability to promote healing in transplant assays, for example post-infarction. While many challenges remain, the development of iPS technology provides a means to generate cells for autologous transplant and for investigating patient-specific disease mechanisms. The development of new techniques to derive cardiac derivatives in vitro from ESC or iPS sources, coupled with novel tissue-engineering approaches and a better understanding of how explanted cells can survive and integrate in host tissue, should have a significant impact on the development of both cell-based and pharmacological therapies for cardiovascular disease. Keywords cardiogenesis; progenitors; cell therapies; iPSCan heart development and disease be studied in vitro?Organogenesis is a complex process that transforms relatively homogenous epithelial germ layers into functioning and highly integrated systems. From a developmental perspective, this can be viewed as a series of cellular transitions, including the commitment of progenitors to tissue-restricted fates, differentiation to express lineage-restricted genetic programs, morphogenesis to form appropriate tissue shapes, and system integration to incorporate the organ into the physiological state of the developing embryo. While the development of any organ system is remarkably complex, cardiogenesis requires extraordinary coordination of regulatory mechanisms in both time and space. In particular, both subtle and dramatic morphogenetic movements transform an initial primordial tube into a complex 3-dimensional organ consisting of septated chambers with distinct identities, a coronary vasculature and mature valves. Integration with both the venous and arterial systems must be timed perfectly 1300 York Ave. LC-709, New York, NY 10021, 212-746-5143, 212-746-8753 (fax), tre2003@med.cornell.edu. Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors...

show abstract

Highly enriched cardiomyocytes from human embryonic stem cells

Cited by 100 publications

References 41 publications

A P19 Cardiac Cell Line as a Model for Evaluating Cardiac Tissue Engineering Biomaterials

A P19 Cardiac Cell Line as a Model for Evaluating Cardiac Tissue Engineering Biomaterials

Transplantation and Tracking of Human-Induced Pluripotent Stem Cells in a Pig Model of Myocardial Infarction

Embryonic stem cells as a model for cardiac development and disease

Contact Info

Product

Resources

About